The two protein components, II and III, of the 4-chlorobenzoate dehalogenase from sp. CBS3 were cloned separately into Component II was obtained on plasmid pCBSII, containing a 3.0 kbp dlll fragment, and component III on plasmid pCBSIIIb, containing a 1.3 kbp I/I fragment. The identities of the two components were confirmed by comparison with the authentic components from sp. CBS3. Both components were expressed constitutively in Neither component alone showed dehalogenating activity. Only in the mixture of crude extracts from both clones was 4-chlorobenzoate dehalogenase detectable. The specific activities in crude extracts were 2.9 mU (mg protein) for component II and 3.5 mU (mg protein) for component III. Expression analysis by minicell experiments revealed a single polypeptide chain of 29 kDa for component II and of 31 kDa for component III.


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